Visualization of Cellular Membranes in 2D and 3D Conditions Using a New Fluorescent Dithienothiophene S,S-Dioxide Derivative.

Cellular membranes play a key role in cell communication with the extracellular environment and neighboring cells. Any changes, including their composition, packing, physicochemical properties and formation of membrane protrusions may affect cells feature. Despite its great importance, tracking membrane changes in living cells is still a challenge. For investigation of processes related to tissue regeneration and cancer metastasis, such as the induction of epithelial-mesenchymal transition, increased cell motility, and blebbing, the possibility to conduct prolonged observation of membrane changes is beneficial, albeit difficult. A particular challenge is conducting this type of research under detachment conditions. In the current manuscript, a new dithienothiophene S,S-dioxide (DTTDO) derivative is presented as an effective dye for staining the membranes of living cells. The synthetic procedures, physicochemical properties, and biological activity of the new compound are presented herein. In addition to the labeling of the membranes in a monolayer culture, its usefulness for visualization of membranes under detachment conditions is also demonstrated. Obtained data have proven that a new DTTDO derivative may be used to stain membranes in various types of experimental procedures, from traditional 2D cell cultures to unanchored conditions. Moreover, due to the specific optical properties, the background signal is reduced and, thus, observation may be performed without washing.

Red-Emitting Dithienothiophene S,S-Dioxide Dyes for Cellular Membrane Staining.

A series of dithienothiophene S,S-dioxide (DTTDO) dyes was designed, synthesized, and investigated for their suitability in fluorescent cell imaging. Synthetized (D-π-A-π-D)-type DTTDO derivatives have molecule lengths close to the thickness of the phospholipid membrane, and they contain on both ends two positively charged or neutral polar groups to increase their solubility in water and to ensure simultaneous interaction with polar groups of the inner and outer part of the cellular membrane. DTTDO derivatives exhibit absorbance and emission maxima in the 517-538 nm and 622-694 nm range, respectively, and a large Stokes shift up to 174 nm. Fluorescence microscopy experiments revealed that these compounds selectively intercalate into cell membranes. Moreover, a cytotoxicity assay conducted on a model human live cells indicates low toxicity of these compounds at the concentrations required for effective staining. With suitable optical properties, low cytotoxicity, and high selectivity against cellular structures, DTTDO derivatives are proven to be attractive dyes for fluorescence-based bioimaging.

Phenylsilane as an effective desulfinylation reagent.

The reduction using phenylsilane in a KOH-catalyzed system was applied successfully to the conversion of sulfinyl-substituted cyclopropylcarboxylates into the corresponding alcohols. The presence of sulfinyl substituents in the α-position to the carboxylate group caused a desulfinylation product formation with full regio- and stereoselectivity, instead of a carbonyl group reduction. Investigations performed on different α-sulfinylcarbonyl compounds revealed that phenylsilane treatment constitutes a regiospecific method for the desulfinylation of a-sulfinylesters; for corresponding ketones the reaction course depends on the character of the carbonyl group.

Highly Asymmetric Cyclopropanation With Two Sulfinyl Auxiliaries as a Way to a Useful Synthetic Intermediate.

The asymmetric cyclopropanation using double sulfinyl auxiliary gave bis-sulfoxide with full diastereoselectivity and very high enantioselectivity. One of the sulfinyl substituents can be removed by sulfinyl metal exchange. Differentiation of sulfinyl substituents made it possible to assign the place of attack of the Grignard reagent.

General route to racemic and enantiomeric carbo- and heterocyclic vinyl sulfoxides via tandem Michael addition/Horner olefination of alpha-phosphorylvinyl sulfoxides.

A new one-pot synthesis of heterocyclic and carbocyclic vinyl sulfoxides has been developed which involves reaction of alpha-phosphorylvinyl sulfoxides with carbonyl compounds bearing oxygen, nitrogen, and carbon nucleophilic centers. Use of optically active alpha-phosphorylvinyl p-tolyl sulfoxides in this tandem Michael addition/Horner olefination reaction leads to the corresponding optically active cyclic sulfoxides. In this way, a variety of optically active chromene, pyrrolizine, chinoline, and cyclopentene sulfoxides have been efficiently prepared.

Stable optically pure phosphino(silyl)carbenes: reagents for highly enantioselective cyclopropanation reactions.

The stability of phosphino(trimethylsilyl)carbenes bearing cyclic diamino substituents on phosphorus is strongly dependent on the steric hindrance of the nitrogen substituents. Phosphinocarbenes 3 and 7, derived from the trans-N,N'-diisopropylcyclohexane-1,2-diamine and N,N'-diisopropyl-1,2-ethanediamine, are not observed; instead the 1,3-diphosphete 4 and a novel six-membered heterocycle 8, which results from the dimerization of 3 and the reaction of 7 with its diazo precursor 6, respectively, have been isolated. In contrast, the phosphino(silyl)carbene 14 derived from N,N'-di-tert-butyl-1,2-ethanediamine has been isolated in high yield. By using the enantiomerically pure (S,S)-, and (R,R)-N,N'-di-tert-butyl-1,2-diphenyl-1,2-ethanediamines, the first optically pure phosphino(sily)carbenes (S,S)-17 and (R,R)-17 have been prepared. They react with methyl acrylate to give the corresponding cyclopropanes (S,S,R,R)-19 and (R,R,S,S)-19 with a total syn diastereoselectivity and an excellent enantioselectivity (de>98 %).